A method for Porphyromonas gingivalis based on recombinase polymerase amplification and lateral flow strip technology.

OBJECTIVE: A practical visual detection method was established to detect Porphyromonas gingivalis (P. gingivalis) by employing a combination of recombinase polymerase amplification and lateral flow strips (RPA-LF) assay, designed for conducting point-of-care testing in clinical settings. METHODS: Primers and probes targeting the P. gingivalis pepO gene were designed. The RPA-LF assay was established by optimising reaction temperature and time, determining the limit of detection (LOD). The specificity of the method was determined by assessing its cross-reactivity with deoxyribonucleic acid from 23 pathogenic bacteria. Finally, the clinical samples from healthy controls (n = 30) and individuals with periodontitis (n = 31) were analysed. The results were compared with those obtained using real-time polymerase chain reaction (PCR). RESULTS: The optimal reaction temperature and time were 39 °C and 12 min. The method exhibited a LOD at 6.40 × 10-4 µg/mL and demonstrated high specificity and sensitivity during cross-reactivity assessment. The RPA-LF assay achieved a P. gingivalis detection rate of 84 % in individuals with periodontitis and 3 % in healthy controls. The results were consistent with those obtained through real-time PCR. CONCLUSION: An RPA-LF assay was developed for detecting P. gingivalis, characterised by its high sensitivity, high specificity, simple operational procedure, and rapid reaction time.

Application of personalized templates in minimally invasive management of coronal dens invaginatus: a report of two cases.

BACKGROUND: Treating the coronal dens invaginatus (CDI) with pulp infection commonly involves the removal of invagination, which increases the risk of perforation and fracture, and compromises the tooth structure. Minimally invasive endodontic management of CDI is highly recommended. This report describes two cases of type II CDI with the application of personalized templates. CASE PRESENTATION: Two cases of type II CDI, affecting the main root canal in a maxillary canine and a lateral incisor, were diagnosed. A guided endodontics (GE) approach was applied. Cone-beam computed tomography and intraoral scans were imported and aligned in a virtual planning software to design debridement routes and templates. The MICRO principle (which involves the aspects of Mechanical (M) debridement, Irrigation (I), Access cavities (C), Rectilinear routes (R), and Obstruction (O)) was proposed for designing optimal debridement routes for future applications. The templates were innovatively personalized and designed to preserve the tooth structure maximally while effectively debriding the root canal. Root canal treatment with supplementary disinfection was then performed. The follow-up of the two patients revealed favorable clinical and radiographic outcomes. CONCLUSIONS: The GE approach could be a feasible method for preserving healthy dental structure while effectively debriding the root canal, thereby achieving successful and minimally invasive endodontic treatment for CDI.

PANoptosis is a compound death in periodontitis: A systematic review of ex vivo and in vivo studies.

OBJECTIVE: The purpose of the systematic review is to verify the presence of PANoptosis in periodontitis based on the published literatures studying cell death in periodontitis. MATERIALS AND METHODS: We conducted a comprehensive review of literature studying the types of cell death in vitro cellular experiments, in vivo rodent studies and clinical studies from three major databases: PubMed, Scopus, and Web of Science. The present systematic review was recorded in the PROSPERO database, under registration number CRD42022383456. RESULTS: In total, 51 articles were included in this study. Our analysis of in vitro cell models revealed that pyroptosis, necroptosis, and apoptosis could be induced by periodontal pathogens in macrophages, fibroblasts, stem cells, and periodontal ligament cells. Furthermore, three types of cell death were detected in in vivo rodent periodontitis models. Clinical studies on human periodontitis tissue specimens and gingival crevicular fluid (GCF) showed that some key proteins related to pyroptosis, necroptosis, and apoptosis were elevated in periodontitis. CONCLUSIONS: Various studies have established similar in vivo and in vitro models with three modes of death detected under the same conditions, revealing complex interactions between different types of cell death pathways in periodontitis and the potential for PANoptosis to occur in periodontitis.

Association between polymorphisms of anti-inflammatory gene alleles and periodontitis risk in a Chinese Han population.

OBJECTIVE: Cytokines that mediate the immune responses are important in the pathogenesis of periodontitis. The genetic polymorphisms of IL-10, TNFAIP3 (A20), and NF-κB1 (p105/p50) and their association with the risk of periodontitis were investigated. METHOD: Venous blood from 102 clinical periodontal healthy participants and 100 patients with periodontitis was collected to genotype the IL-10 (rs1800872), A20 (rs2230926, rs5029937, rs6927127), and NF-κB1 (rs28362491) SNP loci by Sanger technology. Univariable and multivariable logic regression and path analysis model was used to analyze the genotypes and alleles. RESULT: Single-gene mutations in the A20 (rs2230926, rs5029937, rs6927127) and IL-10 (rs1800872) genes were not associated with the risk of periodontitis. NF-κΒ1 (rs28362491) gene influenced periodontitis susceptibility by affecting CAL. The combined effect of A20 and IL-10 was related to the risk of periodontitis (ORa = 0.123-0.151). One site mutated in the A20 (rs2230926, rs5029937, rs6927127) gene or IL-10 (rs1800872) gene reduced the risk of periodontitis. CONCLUSION: Single gene polymorphisms in A20 and IL-10 genes were not associated with the risk of periodontitis. NF-κB1 gene polymorphism indirectly affects susceptibility to periodontitis. The combined effect of anti-inflammatory gene polymorphisms (A20 and IL-10) correlated with the decreased risk of periodontitis. CLINICAL RELEVANCE: This study helps to explore the potential mechanisms underlying the role of anti-inflammatory genes in the progression of periodontal disease and provides a basis for the selection and development of appropriate periodontal treatment strategies based on the genetic profile of the patient.

The efficacy of mouthwashes on oral microorganisms and gingivitis in patients undergoing orthodontic treatment: a systematic review and meta-analysis.

BACKGROUND: Mouthwashes were convenient adjuncts to mechanical cleaning procedures. This review aimed to evaluate the efficacy of mouthwashes on oral microorganisms and gingivitis in orthodontic patients. METHODS: By April 16, 2022, multiple databases and grey literature were searched based on the PICOS strategy. Randomized controlled trials in orthodontic patients evaluating the efficacy of mouthwashes with at least one microbial parameter and/or plaque- and/or gingival inflammation-related index were included. Relevant data were extracted, and the risk of bias was evaluated using Cochrane's tool. Individual mean and standard deviation of the outcomes in mouthwashes and placebos/blank controls were pooled to estimate the weighted mean differences (WMDs) and 95% confidence intervals (95%CIs). Sensitivity analysis, and certainty of evidence were evaluated. RESULTS: Of 1684 articles, 32 studies satisfied the eligibility criteria, and nine were included for meta-analysis. Missing outcome data was the primary source of bias. Compared to blank controls, the short-term application of fluoride mouthwashes significantly reduced the colony counts of Mutans streptococci (MS), while the long-term application may not be effective. Compared to placebos or blank controls, Chlorhexidine mouthwashes significantly reduced the colony counts of multiple microorganisms in the short-term. Compared to placebos or blank controls, herbal mouthwashes showed the inhibitory effect of MS in the short-term, with some results lacking statistical significance. After meta-analysis, significant lower plaque- and gingival inflammation-related indexes were observed in the Chlorhexidine mouthwashes groups [Gingival Index: WMD = -0.45, 95%CI = -0.70 to -0.20 (placebos as control); WMD = -0.54, 95%CI = -0.96 to -0.13 (blank controls); Plaque Index: WMD = -0.70, 95%CI = -1.12 to -0.27 (blank controls)]. Significant lower gingival inflammation-related indexes were observed in the herbal mouthwashes groups [Gingival Index: WMD = -0.20, 95%CI = -0.32 to -0.09 (blank controls)]. CONCLUSIONS: The short-term application of fluoride mouthwashes may reduce the colony counts of cariogenic bacteria, but the long-term effect is not evident. Chlorhexidine may reduce the colony counts of multiple microorganisms in the short-term. Short-term application Chlorhexidine and herbal mouthwashes may effectively reduce plaque- and gingival inflammation-related indexes. However, the risk of bias, inconsistency, and imprecision in the included studies may reduce the certainty of the evidence.

Tilt-angle stimulated Raman projection tomography.

Stimulated Raman projection tomography is a label-free volumetric chemical imaging technology allowing three-dimensional (3D) reconstruction of chemical distribution in a biological sample from the angle-dependent stimulated Raman scattering projection images. However, the projection image acquisition process requires rotating the sample contained in a capillary glass held by a complicated sample rotation stage, limiting the volumetric imaging speed, and inhibiting the study of living samples. Here, we report a tilt-angle stimulated Raman projection tomography (TSPRT) system which acquires angle-dependent projection images by utilizing tilt-angle beams to image the sample from different azimuth angles sequentially. The TSRPT system, which is free of sample rotation, enables rapid scanning of different views by a tailor-designed four-galvo-mirror scanning system. We present the design of the optical system, the theory, and calibration procedure for chemical tomographic reconstruction. 3D vibrational images of polystyrene beads and C. elegans are demonstrated in the C-H vibrational region.

Canonical and noncanonical pyroptosis are both activated in periodontal inflammation and bone resorption.

BACKGROUND AND OBJECTIVE: Pyroptosis has both a caspase-1-dependent canonical pathway and a caspase-4/-5/-11-dependent noncanonical pathway. They play an important role in inflammatory damage and related diseases. Canonical pyroptosis was reported to be involved in periodontitis. However, knowledge of caspase-4/-5/-11-dependent noncanonical pathway involvement remains limited. The aim of this study was to investigate the outcomes of pyroptosis inhibition on periodontitis as well as the possible mechanism, in order to provide a potential target for alleviating periodontitis. METHODS: Human and rat periodontitis tissues were collected for immunohistochemistry (IHC). Micro-computed tomography was used to assess alveolar bone loss in experimental periodontitis model. Pyroptosis-related proteins were tested by western blot. propidium iodide staining and lactate dehydrogenase release were used to verify pyroptosis activation. RNA sequencing was applied to investigate the preliminary mechanism of the reduced periodontal inflammation induced by YVAD-CHO. RESULTS: Both canonical- and noncanonical-related proteins were detected in human and rat periodontitis tissue. The pyroptosis-inhibited group demonstrated less inflammatory response and bone absorption. In vitro, pyroptosis was activated by lipopolysaccharide and inhibited by YVAD-CHO. RNA sequencing demonstrated that the expression of A20 and IκB-ζ was increased and verified by western blot in vitro and IHC in vivo. CONCLUSION: These results suggest that inhibition of pyroptosis-reduced inflammation and alveolar bone resorption in periodontitis.

The correlation of altitude with gingival status among adolescents in western China: a cross-sectional study.

Periodontal disease is common in Chinese adolescents. There is little information about the effect of different altitudes on gingival health. This study aimed to investigate the gingival status at different altitudes and to identify relative factors that affect adolescents' gingival status. A total of 1033 adolescents aged 12-14 years were included in this cross-sectional study in Ganzi (plateau, 1400 m, 2560 m, 3300 m) and Suining (plain, 300 m). Gingival status was assessed by the presence of gingival bleeding on probing (BOP) and dental calculus (DC). Demographic variables, socioeconomic status, dairy habits and oral health-related knowledge, attitudes and behaviors were obtained via questionnaire. Univariate and multivariate binary logistic regression analyses were performed to identify potential relative factors. A total of 64.09% and 77.15% of adolescents had BOP and DC, respectively. The prevalence rates of BOP and DC were higher in the plateau than the plain (P < 0.05). After adjusting for all other factors and interaction terms, residence altitudes of 2560 m [300 m as reference: P < 0.001, odds ratio (OR) = 4.072] and 3300 m (300 m as reference: P = 0.002, OR = 4.053) were significant relative factors of BOP, and an altitude of 2560 m (300 m as reference: P = 0.001, OR = 3.866, 1400 m as reference: P = 0.001, OR = 3.944) was an important relative factor of DC. Gingival bleeding and calculus deposits were common at different altitudes. High altitude was a significant relative factor of gingival bleeding and calculus deposits.

The effects of oral health education regarding periodontal health on non-dental undergraduates in southwestern China-exploring the feasibility of an e-learning course for oral health promotion.

BACKGROUND: The high prevalence of periodontal diseases is an important oral health problem worldwide. It is necessary to increase public knowledge on and influence attitudes towards periodontal diseases in order to prevent them. However, the effect of oral health education (OHE) as a primary preventive method in China is unsatisfactory. The aim of this study is to investigate the feasibility of extending an e-learning course regarding periodontal health by comparing the effects of oral health education regarding periodontal health (OHE-PH) on dental and non-dental students and the effects between a traditional course and an e-learning course among non-dental students at Sichuan University. METHODS: A quasi-experimental study with a pre-test and a post-test was performed. A total of 217 dental students and 134 non-dental students attended a traditional course; 69 non-dental students attended an e-learning course. Before- and after-course questionnaires about periodontal health knowledge, attitudes and behaviours were administered. RESULTS: After the traditional/e-learning course, the knowledge of both dental and non-dental students about periodontal diseases and self-reported behaviours for gingival bleeding and oral care improved. The non-dental students reached or surpassed the level of dental students before the course. The non-dental students taking the e-learning course performed better in some areas than those taking the traditional course. CONCLUSIONS: OHE-PH was effective for dental and non-dental students. The e-learning course on OHE-PH was sufficient for improving knowledge and self-reported behaviours among non-dental undergraduates and was even better than the traditional course in some areas. The e-learning course may be an effective method for periodontal health education and oral health promotion among undergraduates.

A predictive nomogram: a cross-sectional study on a simple-to-use model for screening 12-year-old children for severe caries in middle schools.

BACKGROUND: A nomogram is a tool that transforms complex regression equations into simple and visual graphs and enables clinicians and patients to conveniently compute output probabilities without needing medical knowledge and complex formulas. The aim of this study was to develop and validate a predictive nomogram to screen for severe caries among 12-year-old children based on risk factors in Sichuan Province, China. METHODS: A cross-sectional study of 4573 12-year-olds was conducted up to May 2016 in middle schools from three districts and three counties in Sichuan Province, China. All the children underwent oral examinations and completed questionnaires to assess general information, oral impacts on daily performance, dietary habits, subjective health conditions, history of dental trauma, frequency of toothache, dental visits, and knowledge, attitudes, and behaviours toward oral hygiene. Univariate analysis and multivariate logistic regression analysis were used to determine which variables were significantly associated with severe caries (operationalized as DMFT ≥ 3). A nomogram was developed and validated by using the 'rms' package and two cross-validation methods. RESULTS: Severe caries was found in 537 of the 4573 children (11.74%). Multivariate logistic regression analysis revealed that the following variables predicted a higher risk of severe caries: 'female' [odds ratio (OR) = 1.985, 95% confidence interval (95% CI): 1.63-2.411], 'urban' (OR = 2.389, 95% CI: 1.96-2.91), 'non-only child' (OR = 1.317, 95% CI: 1.07-1.625), 'very poor self-assessment of oral health status' (OR = 2.157, 95% CI: 1.34-3.467) and 'visited a dentist less than 6 months' (OR = 1.861, 95% CI: 1.38-2.505). Multivariate logistic regression analysis also indicated that the following variables predicted a lower risk of severe caries: 'middle level of urbanization' (OR = 0.395, 95% CI: 0.32-0.495) and 'high level of urbanization' (OR = 0.466, 95% CI: 0.37-0.596). Both the fivefold and leave-one-out cross-validation methods indicated that the nomogram model built by these 6 variables displayed good disease recognition ability. CONCLUSIONS: The nomogram was a simple-to-use model to screen children for severe caries. This model was found to facilitate non-dental professionals in assessing risk values without oral examinations and making referrals to dental professionals.

Comparison of oral health behaviour between dental and non-dental undergraduates in a university in southwestern China--exploring the future priority for oral health education.

BACKGROUND: Based on a national survey in 2015, people's oral health behaviour (OHB) has not kept up with the pace of knowledge and attitudes in China after decades of oral health education (OHE). Thus, we need to improve OHE to strengthen people's OHB. Undergraduates are regarded as the best candidates for the improvement of OHE. The objective of this study is to determine undergraduates' oral health status and existing problems in OHB by comparing dental and non-dental students at Sichuan University. We hope to provide some suggestions for future OHE to improve people's OHB. METHODS: A quasi-experimental study designed with a pre-test and post-test group was conducted. A total of 217 dental students and 135 non-dental students were enrolled. They were administered an OHE course focused on OHB. A survey about oral health behaviour and knowledge was conducted before and after the course. RESULTS: According to the pre-course survey, dental students surpassed non-dental students in terms of toothbrushing frequency, method, and time, but unfortunately, flossing was overlooked by all the students. After the course, both dental and non-dental students showed strong willingness to improve their OHB. More non-dental students than dental students were willing to use toothpicks and Chinese herbal toothpaste before and after the course. CONCLUSIONS: OHE focused on behaviour has a positive effect on university students. Future OHE and interventions should focus on flossing, toothbrushing methods, toothpicks, Chinese herbal toothpaste and modifications to adopt new media.

Gingival bleeding and calculus among 12-year-old Chinese adolescents: a multilevel analysis.

BACKGROUND: Gingivitis is a common oral health problem, and untreated gingivitis can progress to periodontitis. The objectives of this study were to (1) explore associated factors of gingival bleeding and calculus among 12-year-old adolescents; (2) find predictive models for gingivitis management. METHODS: Four thousand five hundred twenty-five subjects aged 12 in Sichuan Province were investigated. The questionnaire and clinical examination were applied in schools, and two-level logistic regression models were constructed to interpret the effect of individual and contextual factors on Chinese adolescents' gingival bleeding and calculus. RESULTS: 46.63% (95%CI: 40.71, 51.60) and 66.94% (95%CI: 56.85, 67.45) of the subjects presented gingival bleeding and calculus, respectively. For the gingival bleeding cases, the model showed the significant associated indicators were hukou (OR = 0.61, 95% CI: 0.52-0.72), family size (OR = 1.41, 95% CI: 1.19-1.68), parental educational level (father: OR = 0.53, 95% CI: 0.45-0.63; mother: OR = 0.71, 95% CI: 0.59-0.86), tooth-brushing frequency (OR = 0.35, 95% CI: 0.26-0.48), dental floss use (OR = 0.58, 95% CI: 0.41-0.83), sugar-containing drink consumption (OR = 2.11, 95% CI: 1.80-2.49), and dental visit (OR = 1.44, 95% CI: 1.19-1.74). It also confirmed that gender (OR = 1.32, 95% CI: 1.13-1.54), hukou (OR = 0.69, 95% CI: 0.59-0.82), family size (OR = 1.34, 95% CI: 1.12-1.59), parental educational level (father: OR = 0.46, 95% CI: 0.39-0.54; mother: OR = 0.65, 95% CI: 0.59-0.82), tooth-brushing frequency (OR = 0.57, 95% CI: 0.42-0.78), dental floss use (OR = 0.66, 95% CI: 0.48-0.90) and sugar-containing drink consumption (OR = 1.30, 95% CI: 1.11-1.53) were associated factors for dental calculus. CONCLUSIONS: Gingival bleeding and calculus were common in western Chinese adolescents. Socio-demographic factors including gender, hukou and family factors are strong determinants of gingival health in Chinese adolescents. In addition, health-related lifestyle behaviors such as healthy diet, good hygiene care and more dental visits are good predictors of better gingival status.

Application of a three-session-procedure based on experiential learning in a tooth brushing course for Chinese dental students.

BACKGROUND: Chinese dentists are obliged to provide reliable guidance to patients about tooth brushing. However, attitudes and behaviours of Chinese dental students regarding oral health have been insufficient. Traditionally, dental students were taught how to brush, but not how to evaluate tooth brushing. Here, we applied a three-session-procedure (TSP) based on experiential learning (EL) in a tooth brushing course for Chinese dental students. The aim was to improve dental students' knowledge, practice of tooth brushing and self-evaluation, which may help cultivate their ability to conduct evaluation for friends, relatives and future patients. METHODS: A quasi-experimental study design with a pre-test and post-test group was applied. A total of 176 students were enrolled in the TSP, which included a 1.5-h lecture course, a 3-h practice course for evaluation and comparison of manual and electric tooth brushing, and an after-class experience report. A survey including a knowledge test and a questionnaire on tooth brushing habits and opinions about the TSP was conducted 2 weeks later. The data about tooth brushing and the survey were collected and analysed. RESULTS: Manual and electric tooth brush demonstrated almost equal overall efficiency in reducing plaque. However, for some students, either manual or electric tooth brushing was more suitable. Thus, it is advisable to estimate the exact differences in the efficiency of tooth brushing methods for each individual. The survey showed that tooth brushing by dental students significantly improved after TSP. The students could make self-evaluation and proper recommendations to family members and friends. The TSP was helpful in interpreting and evaluating manual and electric tooth brushing methods. CONCLUSIONS: A TSP course improved dental students' knowledge, practice of tooth brushing and self-evaluation. In conclusion, a TSP based on EL is an effective and well-organized method of education on tooth brushing for Chinese dental students.

The extent of pyroptosis varies in different stages of apical periodontitis.

Apical periodontitis (AP) is an inflammation affecting the periapical region of tooth root. Microbial pathogens activate inflammasomes and promote the production of pro-inflammatory cytokines. Caspase-1-mediated pyroptosis is a possible mechanism involved in the initiation and progression of AP. The purpose of this study was to evaluate the role of caspase-1 and pyroptosis on AP at different stages. Human periapical inflammatory tissue was collected to study chronic AP stage. Human periodontal ligament fibroblasts (hPDLFs) were stimulated with lipopolysaccharide in vitro for 24h to simulate early AP stage. Experimental AP rat model was established to study acute AP stage from 0d to 28d. The results showed that NLRP3, cleaved caspase-1 and Interleukin (IL)-1ß were enhanced in all AP stages. Caspase-1 activation was detectable in most cells. However, the level of pyroptosis was in accordance with the degree of AP inflammation. Early and chronic AP showed a comparable hemostasis state, with pyroptosis remaining in a reduced level. On the contrary, extensive pyroptosis accelerated inflammation and induced cell death in acute AP. VX765, a caspase-1 inhibitor, was used in an experimental AP rat model. The results showed that VX765 suppressed bone loss, suggesting a role of pyroptosis on bone resorption in acute AP. VX765 also inhibited the expressions of IL-1ß, Monocyte chemoattractant protein-1 (MCP-1), IL-6 and IL-8 in vitro, thus decreased inflammatory responses during AP. In conclusion, caspase-1 and pyroptosis contributed to AP inflammation and lesion and pyroptosis extent was in line with AP progression.

Distinct oral-associated gastric microbiota and Helicobacter pylori communities for spatial microbial heterogeneity in gastric cancer.

The gastric microbial community plays a fundamental role in gastric cancer (GC), and the two main anatomical subtypes of GC, non-cardia and cardia GC, are associated with different risk factors (Helicobacter pylori for non-cardia GC). To decipher the different microbial spatial communities of GC, we performed a multicenter retrospective analysis to characterize the gastric microbiota in 223 GC patients, including H. pylori-positive or -negative patients, with tumors and paired adjacent normal tissues, using third-generation sequencing. In the independent validation cohort, both dental plaque and GC tumoral tissue samples were collected and sequenced. The prevalence of H. pylori and oral-associated bacteria was verified using fluorescence in situ hybridization (FISH) assays in GC tumoral tissues and matched nontumoral tissues. We found that the vertical distribution of the gastric microbiota, at the upper, middle, and lower third sites of GC, was likely an important factor causing microbial diversity in GC tumor tissues. The oral-associated microbiota cluster, which included Veillonella parvula, Streptococcus oralis, and Prevotella intermedia, was more abundant in the upper third of the GC. However, H. pylori was more abundant in the lower third of the GC and exhibited a significantly high degree of microbial correlation. The oral-associated microbiota module was co-exclusive with H. pylori in the lower third site of the GC tumoral tissue. Importantly, H. pylori-negative GC patients with oral-associated gastric microbiota showed worse overall survival, while the increase in microbial abundance in H. pylori-positive GC patients showed no difference in overall survival. The prevalence of V. parvula in both the dental plaque and GC tissue samples was concordant in the independent validation phase. We showed that the oral-associated species V. parvula and S. oralis were correlated with overall survival. Our study highlights the roles of the oral-associated microbiota in the upper third of the GC. In addition, oral-associated species may serve as noninvasive screening tools for the management of GC and an independent prognostic factor for H. pylori-negative GCs. IMPORTANCE: Our study highlights the roles of the oral-associated microbiota in the upper third of gastric cancer (GC).We showed that the oral-associated species Veillonella parvula and Streptococcus oralis were correlated with overall survival. In addition, oral-associated species may serve as noninvasive screening tools for the management of GC and an independent prognostic factor for Helicobacter pylori-negative GCs.

Metabolic shift and the effect of mitochondrial respiration on the osteogenic differentiation of dental pulp stem cells.

Background: Metabolism shifts from glycolysis to mitochondrial oxidative phosphorylation are vital during the differentiation of stem cells. Mitochondria have a direct function in differentiation. However, the metabolic shift and the effect of mitochondria in regulating the osteogenic differentiation of human dental pulp stem cells (hDPSCs) remain unclear. Methods: Human dental pulp stem cells were collected from five healthy donors. Osteogenic differentiation was induced by osteogenic induction medium. The activities of alkaline phosphatase, hexokinase, pyruvate kinase, and lactate dehydrogenase were analyzed by enzymatic activity kits. The extracellular acidification rate and the mitochondrial oxygen consumption rate were measured. The mRNA levels of COL-1, ALP, TFAM, and NRF1 were analyzed. The protein levels of p-AMPK and AMPK were detected by western blotting. Results: Glycolysis decreased after a slight increase, while mitochondrial oxidative phosphorylation continued to increase when cells grew in osteogenic induction medium. Therefore, the metabolism of differentiating cells switched to mitochondrial respiration. Next, inhibiting mitochondrial respiration with carbonyl cyanide-chlorophenylhydrazone, a mitochondrial uncoupler inhibited hDPSCs differentiation with less ALP activity and decreased ALP and COL-1 mRNA expression. Furthermore, mitochondrial uncoupling led to AMPK activation. 5-Aminoimidazole-4-carboxamide ribonucleotide, an AMPK activator, simulated the effect of mitochondrial uncoupling by inhibiting osteogenic differentiation, mitochondrial biogenesis, and mitochondrial morphology. Mitochondrial uncoupling and activation of AMPK depressed mitochondrial oxidative phosphorylation and inhibited differentiation, suggesting that they may serve as regulators to halt osteogenic differentiation from impaired mitochondrial oxidative phosphorylation.

The Relationship Between Periodontal Disease and Breast Cancer: From Basic Mechanism to Clinical Management and Prevention.

PURPOSE: Periodontal disease is potentially related to certain kinds of cancer. This review aimed to summarize the relationship between periodontal disease and breast cancer, providing some strategies for the clinical treatment and periodontal health care of breast cancer patients. MATERIALS AND METHODS: Systematic reviews, randomised controlled trials, prospective and retrospective clinical studies, case series and reports were collected using search terms entered into the PubMed, Google Scholar and JSTOR databases. RESULTS: Research has provided some evidence that periodontal disease is related to the occurrence and development of breast cancer. Periodontal disease and breast cancer have some common pathogenic factors. Periodontal disease may affect the initiation and development of breast cancer involving microorganisms and inflammation. Periodontal health is affected by radiotherapy, chemotherapy, and endocrine therapy for breast cancer. CONCLUSIONS: Periodontal therapy for breast cancer patients should be performed differently according to the stage of cancer treatment. Adjuvant endocrine treatment (e.g. bisphosphonates) has a great impact on oral treatment. Periodontal therapy contributes to the primary prevention of breast cancer. Periodontal health care of breast cancer patients is worthy of clinician attention.

The Efficacy and Safety of Oral Irrigator on the Control of Dental Plaque and Gingivitis: A Randomized, Single-Blind, Parallel-Group Clinical Trial.

BACKGROUND: To evaluate the efficacy and safety of oral irrigator (OI) in controlling dental plaque and gingivitis. METHODS: Ninety participants diagnosed with gingivitis were randomly assigned to two groups, given a toothbrush combined with OI (WaterPik®) (test) or a toothbrush alone (control). The Turesky-Modified Quigley-Hein Plaque Index (T-QH), Modified Gingival Index (MGI), Bleeding Index (BI), and percentage of sites with bleeding on probing (BOP%) were evaluated at baseline, 4 weeks, 8 weeks, and 12 weeks. The full analysis set (FAS) and per-protocol set (PPS) were analyzed. Adverse events were recorded through electronic diaries and examinations. RESULTS: Of the 90 participants, the efficacy was assessed in the following numbers (FAS/PPS): test (45/33) and control (43/38). Compared with the control, MGI, BI, and BOP% were significantly lower in the test group after 4 weeks (4 weeks: p = 0.017, p = 0.001, and p = 0.001, respectively; 8 weeks and 12 weeks: p < 0.001 for all, FAS); T-QH was significantly lower after 8 weeks (8 weeks: p = 0.033; 12 weeks: p = 0.006, FAS). Transient gingival bleeding may be associated with OI. Self-reported pain and dentin hypersensitivity symptoms were similar between groups. CONCLUSIONS: As adjuncts to toothbrushing, OI demonstrated significantly better efficacy in controlling dental plaque and gingival inflammation with no substantial safety hazards.

Science Popularization Education regarding Oral Health-General Health for Nonmedical Undergraduates Applying a SPOC Teaching Model.

Objective: To see how effective a blended teaching model based on a small private online course (SPOC) is in a science popularization education course on oral health-general health (OHGH). Methods: The SPOC blended teaching model was created using an elective classroom course "Oral Prophylaxis and Hygiene" in conjunction with an online learning course called "Preventive Dentistry" from the China University massive open online course (MOOC) for the science popularization education on OHGH. Students' evaluations and teaching efficacy of this science popularization education course were tested using pre- and postcourse questionnaires. Results: In all, 105 valid questionnaires were returned. Before the course, 95.2% of the students expressed an interest in learning more knowledge on oral disease and OHGH. When compared to those of the precourse, students' knowledge of oral diseases and OHGH was significantly higher (P < 0.0001) and the associated practice after science popularization education was much increased (P < 0.0001 or P = 0.0005), except for root canal therapy (P = 0.3886). The scores of students on the scientific popularization task also improved when compared to those of the previous classroom-only teaching (P < 0.0001). In the postcourse questionnaire, students rated the SPOC teaching mode significantly higher than both online learning and classroom teaching alone (P < 0.0001; P = 0.0117); the SPOC blended teaching was judged as more suitable for science popularization education (P < 0.0001). Conclusion: The application of the SPOC teaching mode for the science popularization education course on OHGH to nonmedical undergraduates has better teaching outcomes and is more likely to be accepted by college students.

Double-Edged Sword Effect of Pyroptosis: The Role of Caspase-1/-4/-5/-11 in Different Levels of Apical Periodontitis.

The study was to investigate the effect of canonical and noncanonical pyroptosis in apical periodontitis. Proteins' profiles of human apical periodontitis tissue were analyzed by label-free proteomics. Immunofluorescence was used to detect proteins related to pyroptosis in human apical periodontitis tissues and experimental apical periodontitis models. A dual experimental apical periodontitis model with both smaller (mandible) and larger (maxilla) bone lesions was established. THP-1-derived macrophages were stimulated with P. gingivalis lipopolysaccharide in vitro with or without the caspase-1/-4/-5 inhibitor Ac-FTDL-CMK. Propidium iodide staining, lactic dehydrogenase release and Western blot were applied to evaluate cell death and the protein expression. Caspase-1/-4/-5 were expressed in human apical periodontitis tissues. Caspase-1/-11 were involved in bone loss in experimental apical periodontitis. Caspase-1/-11 inhibitors reduced bone loss in larger lesions (maxilla) but accelerated bone loss in smaller lesions (mandible). Caspase-1/-4/-5 inhibitors also showed double-edged sword effects on propidium iodide staining and lactic dehydrogenase release in vitro. The expression of cleaved-caspase-1/-4/-5, mature interluekin-1ß and gasdermin D N-terminal domain increased in THP-1-derived macrophages after lipopolysaccharide stimulation but decreased after treatment with Ac-FTDL-CMK. Pyroptosis contributed to apical periodontitis and excited a double-edged sword effect in inducing bone loss in vivo and cell death in vitro.